IL157274A - Human monoclonal antibodies to cd89, antigen binding portions thereof, hybridomas and transfectomas producing them, transgenic cells expressing them, bispecific, multispecific, molecule conjugates, immunotoxins and compositions comprising them, methods of production thereof and methods of detection thereof - Google Patents

Human monoclonal antibodies to cd89, antigen binding portions thereof, hybridomas and transfectomas producing them, transgenic cells expressing them, bispecific, multispecific, molecule conjugates, immunotoxins and compositions comprising them, methods of production thereof and methods of detection thereof

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Publication number
IL157274A
IL157274A IL157274A IL15727403A IL157274A IL 157274 A IL157274 A IL 157274A IL 157274 A IL157274 A IL 157274A IL 15727403 A IL15727403 A IL 15727403A IL 157274 A IL157274 A IL 157274A
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IL
Israel
Prior art keywords
human
antibody
amino acid
variable region
antigen binding
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IL157274A
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Hebrew (he)
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Medarex Inc
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Publication date
Application filed by Medarex Inc filed Critical Medarex Inc
Publication of IL157274A publication Critical patent/IL157274A/en

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    • C07K16/28—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
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    • C07K16/2803—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily
    • C07K16/283—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily against Fc-receptors, e.g. CD16, CD32, CD64
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    • G01N2333/70503—Immunoglobulin superfamily, e.g. VCAMs, PECAM, LFA-3
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Claims (57)

- 69 - 157274/2 CLAIMS:
1. An isolated human monoclonal antibody, or antigen binding portion thereof, which binds to human CD89, comprising a human heavy chain variable region comprising CRD1, CDR2, and CD 3 sequences and a human light chain variable region comprising CDR1, CDR2, and CDR3 sequences, wherein (a) the human heavy chain variable region CDR3 sequence comprises an amino acid sequence selected from the group consisting of amino acid residues 99-108 of SEQ ID NO:2, amino acid residues 99-108 of SEQ ID NO:6, and conservative sequence modifications thereof; or (b) the human light chain variable region CDR3 sequence comprises an amino acid sequence selected from the group consisting of amino acid residues 89-97 of SEQ ID NO:4, amino acid residues 90-99 of SEQ ID NO: 8, and conservative sequence modifications thereof.
2. The antibody, or antigen binding portion thereof, of claim 1, wherein the human heavy chain variable region CDR2 sequence comprises an amino acid sequence selected from the group consisting of amino acid residues 50-66 of SEQ ID NO:2, amino acid residues 50-66 of SEQ ID NO:6, and conservative sequence modifications thereof; and the human light variable region CDR2 sequence comprises an amino acid sequence selected from the group consisting of amino acid residues 50-56 of SEQ ID NO:4, amino acid residues 51-57 of SEQ ID NO:8, and conservative sequence modifications thereof.
3. The .antibody, or antigen binding portion thereof, of claim 1, wherein the human heavy chain variable region CDR1 sequence comprises an amino acid sequence selected from the group consisting of amino acid residues 30-35 of SEQ ID NO:2, amino acid residues 31-35 of SEQ ID NO:6, and conservative sequence modifications thereof; and the human light variable region CDR1 sequence comprises an amino acid sequence selected from the group consisting of amino acid residues 24-34 of SEQ ID NO:4, amino acid residues 24-35 of SEQ ID NO:8, and conservative sequence modifications thereof.
4. An isolated human monoclonal antibody, or antigen binding portion thereof, comprising (i) a heavy chain variable region comprising CDR1, CDR2, and - 70 - 157274/2 CDR3 sequences comprising amino acid residues 30-35, amino acid residues 50-66, and amino acid residues 99-108 of SEQ ID NO:2, respectively; and (ii) a light chain variable region comprising CDR1, CDR2, and CDR3 sequences comprising amino acid residues 24-34, amino acid residues 50-56, and amino acid residues of 89-97 of SEQ ID NO:4, respectively, wherein the antibody, or antigen binding portion thereof, binds to human CD89.
5. An isolated human monoclonal antibody, or antigen binding portion thereof, comprising (i) a heavy chain variable region comprising CDR1, CDR2, and CDR3 sequences comprising amino acid residues 31-35, amino acid residues 50-66, and amino acid residues 99-108 of SEQ ID NO:6, respectively; and (ii) a light chain variable region comprising CDR1, CDR2, and CDR3 sequences comprising amino acid residues 24-35, amino acid residues 51-57, and amino acid residues of 90-99 of SEQ ID NO: 8, respectively, wherein the antibody, or antigen binding portion thereof, binds to human CD89.
6. An isolated human monoclonal antibody, or antigen binding portion thereof, comprising a human heavy chain variable region and a human light chain variable region, wherein: (a) the human heavy chain variable region comprises an amino acid sequence that is at least 90% homologous to the amino acid sequence of SEQ ID NO:2; (b) the human light chain variable region comprises an amino acid sequence that is at least 90% homologous to the amino acid sequence of SEQ ID NO:4; and (c) the antibody, or antigen binding portion thereof, binds to human CD89.
7. An isolated human monoclonal antibody, or antigen binding portion thereof, comprising a human heavy chain variable region and a human light chain variable region, wherein: (a) the human heavy chain variable region comprises an amino acid sequence that is at least 90% homologous to the amino acid sequence of SEQ ID NO:6; - 71 - 157274/2 (b) the human light chain variable region comprises an amino acid sequence that is at least 90% homologous to the amino acid sequence of SEQ ID NO:8; and (c) the antibody, or antigen binding portion thereof, binds to human CD89.
8. An isolated human monoclonal antibody, or antigen binding portion thereof, comprising human heavy chain and human light chain variable regions comprising the amino acid sequences shown in SEQ ID NO:2 and DEQ ID NO:4, respectively, wherein the antibody, or antigen binding portion thereof, binds to human CD89.
9. An isolated human monoclonal antibody, or antigen binding portion thereof, comprising human heavy chain and human light chain variable regions comprising the amino acid sequences shown in SEQ ID NO:6 and DEQ ID NO:8, respectively, wherein the antibody, or antigen binding portion thereof, binds to human CD89.
10. The isolated human monoclonal antibody, or antigen binding portion thereof, comprising: (a) a heavy chain variable region derived from a human germline 3- 30.3 VH gene; or (b) a heavy chain variable region derived from a human germline VKL18 or VKA27 gene; wherein the human antibody binds human CD89.
11. The isolated human monoclonal antibody, or antigen binding portion thereof, comprising: (a) a heavy chain variable region derived from a human germline 3- 30.3 VH gene; or (b) a heavy chain variable region derived from a human germline VRA27 gene; wherein the human antibody binds human CD89.
12. The isolated human monoclonal antibody, or antigen binding portion thereof, comprising: - 72 - 157274/2 (a) a heavy chain variable region derived from a human germline 3- 30.3 VH gene; or (b) a heavy chain variable region derived from a human germline Vj L18 or VKA27 gene; wherein the human antibody does not activate complement.
13. The antibody of claim 10 or 12, wherein the light chain variable region is derived from a human germline V L18 gene.
14. The antibody of any one of claims 10 to 12, wherein the light chain variable region is derived from a human germline VKA27 gene.
15. The antibody of any one of claims 1 to 14, wherein the antibody is a Fab fragment or a single chain antibody.
16. The antibody of any one of claims 1 to 15, wherein the antibody binds to 8 1 human CD 89 with an affinity constant of at least 10 M" .
17. The antibody of any one of claims 1 to 16, wherein the antibody binds to human CD89 with an affinity constant of at least 109 M"1.
18. The antibody of any one of claims 1 to 12, wherein the antibody heavy chain is an IgGl heavy chain and the antibody light chain is a kappa light chain.
19. A hybridoma comprising a B cell obtained from a transgenic nonhuman animal having a genome comprising a human heavy chain transgene and a light chain transgene fused to an immortalized cell, wherein the hybridoma produces a detectable amount of the antibody, or antigen binding portion thereof, of any one of claims 1 to 18.
20. A transfectoma comprising nucleic acids encoding a human heavy chain and a human light chain, wherein the transfectoma produces a detectable amount of the antibody, or antigen binding portion thereof, of any one of claims 1 to 18.
21. A transgenic cell which expresses the antibody, or antigen binding portion thereof, of any one of claims 1 to 18, wherein the transgenic cell has a genome comprising a human heavy chain transgene or transchromosome and a human light chain transgene. - 73 - 157274/2
22. A method of producing the antibody, or antigen binding portion thereof, of any one of claims 1 to 18, comprising: immunizing a transgenic nonhuman animal having a genome comprising a human heavy chain transgene and a human light chain transgene with human CD 89 or a cell expressing human CD89, such that antibodies are produced by B cells of the animal; isolating B cells of the animal; fusing the B cells with myeloma cells to form immortal, hybridoma cells that secrete human monoclonal antibodies specific for CD89; and isolating the human monoclonal antibodies specific for CD 89 from the culture supernatant of the hybridoma.
23. A bispecific or multispecific molecule comprising the antibody, or antigen binding portion thereof, of any one of claims 1 to 18 and a portion which binds to a target antigen other than CD89.
24. The bispecific or multispecific molecule of claim 23, wherein the antibody is an Fab fragment or a single chain antibody.
25. The bispecific or multispecific molecule of claim 23, wherein the target antigen is a tumor antigen.
26. The bispecific or multispecific molecule of claim 23, wherein the portion that binds to the target antigen comprises an antibody or a tumor ligand.
27. The bispecific or multispecific molecule of claim 23 comprising a fusion protein.
28. The bispecific or multispecific molecule of claim 23 comprising a chemically linked conjugate.
29. The bispecific or multispecific molecule of claim 23 which induces lysis (ADCC) of a cell expressing the target antigen in the presence of effector cells expressing CD89.
30. The bispecific or multispecific molecule of claim 23, wherein the target antigen is selected from the group consisting of carcinoembryonic antigen (CEA), - 74 - 157274/2 gastrin- releasing peptide receptor antigen (GRP), mucine antigens, epidermal growth factor receptor (EGF-R), BER2/neu, HER3, HER4, CD20, CD30, MAGE antigens, SART antigens, MUC1 antigen, c-erb-2 antigen and TAG 72.
31. The bispecific or multispecific molecule of claim 26, wherein the tumor ligand is selected from the group consisting of epidermal growth factor (EGF), bombesin, gastrin-releasing peptide (GRP), litorin, neuromedin B, or neuromedin C.
32. The bispecific or multispecific molecule of claim 26, wherein the tumor ligand is epidermal growth factor (EGF).
33. A molecule conjugate comprising the human antibody, or antigen binding portion thereof, of any one of claims 1 to 18 linked to an antigen.
34. The molecular conjugate of claim 33, wherein the antibody is an Fab fragment or a single chain antibody.
35. The molecular conjugate of claim 33 comprising a fusion protein.
36. The molecular conjugate of claim 33 comprising a chemically linked conjugate.
37. The molecular conjugate of claim 33, wherein the antigen is selected from the group consisting of carcinoembryonic antigen (CEA), gastrin-releasing peptide receptor antigen (GRP), mucine antigens, epidermal growth factor receptor (EGF-R), HER2/neu, HER3, HER4, CD20, CD30, MAGE antigens, SART antigens, MUC1 antigen, c-erb-2 antigen and TAG 72.
38. A composition comprising the antibody, or antigen binding portion thereof, of any one of claims 1 to 18 and a pharmaceutically acceptable carrier.
39. A composition comprising ah isolated human monoclonal antibody, or antigen binding portion thereof, comprising: (a) a heavy chain variable region derived from a human germline 3- 30.3 VH gene; or (b) a heavy chain variable region derived from a human germline VKL18 or VKA27 gene; wherein the human antibody binds human CD89. - 75 - 157274/2
40. A composition comprising the bispecific or multispecific molecule of any one of claims 23 and 32, and a pharmaceutically acceptable carrier.
41. A composition comprising the molecular conjugate of any one of claims 33 to 37 and a pharmaceutically acceptable carrier.
42. A composition comprising a combination of two or more antibodies, or antigen binding portion thereof, any one of claims 1 to 18, wherein each of said antibodies binds to a distinct epitope of human CD89.
43. The composition of claim 38 further comprising a cytotoxic agent.
44. An immunotoxin comprising the antibody, or antigen binding portion thereof, of any one of claims 1 to 18 linked to a cytotoxic agent.
45. A method of detecting the presence of CD89 or a cell expressing CD89 in a sample comprising: contacting the sample with the antibody of any one of claims 1 to 18 under conditions that allow for formation of a complex between the antibody and CD89; and detecting the formation of the complex.
46. An expression vector comprising a nucleotide sequence encoding the variable region of the heavy chain, light chain, or both heavy and light chains of a human monoclonal antibody which binds to human CD89, wherein the heavy chain nucleotide sequence is selected from the group consisting of SEQ ID NO:l and SEQ ID NO: 5 and the light chain nucleotide sequence is selected from the group consisting of SEQ ID NO:3 and SEQ ID NO:7.
47. An isolated human monoclonal antibody, or antigen binding portion thereof, substantially as herein described with reference to any one of the embodiments of the invention illustrated in the accompanying drawings and/or examples.
48. A hybridoma according to claim 19, substantially as herein described with reference to any one of the embodiments of the invention illustrated in the accompanying drawings and/or examples. - 76 - 157274/2
49. A transfectoma according to claim 20, substantially as herein described with reference to any one of the embodiments of the invention illustrated in the accompanying drawings and/or examples.
50. A transgenic cell according to claim 21, substantially as herein described with reference to any one of the embodiments of the invention illustrated in the accompanying drawings and/or examples.
51. A method according to claim 22, substantially as herein described with reference to any one of the embodiments of the invention illustrated in the accompanying drawings and/or examples.
52. A bispecific or multispecific molecule according to claim 23, substantially as herein described with reference to any one of the embodiments of the invention illustrated in the accompanying drawings and/or examples.
53. A molecular conjugate according to claim 33, substantially as herein described with reference to any one of the embodiments of the invention illustrated in the accompanying drawings and/or examples.
54. A composition according to any one of claims 38 to 42, substantially as herein described with reference to any one of the embodiments of the invention illustrated in the accompanying drawings and/or examples.
55. An immunotoxin, substantially as herein described with reference to any one of the embodiments of the invention illustrated in the accompanying drawings and/or examples.
56. A method of detecting the presence of CD89 or a cell expressing CD89 in a sample, substantially as herein described with reference to any one of the embodiments of the invention illustrated in the accompanying drawings and/or examples.
57. An expression vector comprising a nucleotide sequence encoding the variable region of the heavy chain, light chain, or both heavy and light chains of a human monoclonal antibody which binds to human CD89, substantially as herein - 77 - 157274/2 described with reference to any one of the embodiments of the invention illustrated in the accompanying drawings and/or examples. For the Applicants, REINHOLD COHN AN¾ PARTNERS By:
IL157274A 2001-02-12 2003-08-06 Human monoclonal antibodies to cd89, antigen binding portions thereof, hybridomas and transfectomas producing them, transgenic cells expressing them, bispecific, multispecific, molecule conjugates, immunotoxins and compositions comprising them, methods of production thereof and methods of detection thereof IL157274A (en)

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